Richter H, Ennis H L
Arch Biochem Biophys. 1985 Oct;242(1):16-22. doi: 10.1016/0003-9861(85)90474-6.
A repetitive DNA sequence was isolated from a Dictyostelium discoideum genomic plasmid library of BglII-digested DNA ligated to the BamHI site in pBR322. This clone, called pBS582, hybridized to a large number of phage lambda Dictyostelium genomic clones. Southern blot analysis indicated that pBS582 DNA hybridized to many differently sized genomic DNA fragments generated by digestion with Eco RI, AvaI, or HindIII. Restriction maps of pBS582 and five genomic clones showed that the flanking regions of each of the genomic clones were different. These findings indicate that the sequence specific to pBS582 is scattered throughout the Dictyostelium genome and is reiterated approximately 100 times in the haploid genome. Northern blot analysis revealed that RNA which hybridized to pBS582 DNA was present during all stages of growth and development and did not seem to be developmentally regulated. Southern blot analysis of DNAs from other slime molds (D. giganteum, D. purpureum, and Polysphondylium violaceum) were performed to determine whether the pBS582 sequence was present in other species of slime molds. Hybridization of pBS582 was observed to DNA from the two Dictyostelium species but not to Polysphondylium. It may thus be possible to use hybridization of specific sequences as a biochemical tool to study the relatedness of different slime mold species and their molecular taxonomy.
从用BglII消化并连接到pBR322的BamHI位点的盘基网柄菌基因组质粒文库中分离出一个重复DNA序列。这个名为pBS582的克隆与大量噬菌体λ盘基网柄菌基因组克隆杂交。Southern印迹分析表明,pBS582 DNA与用Eco RI、AvaI或HindIII消化产生的许多不同大小的基因组DNA片段杂交。pBS582和五个基因组克隆的限制性图谱显示,每个基因组克隆的侧翼区域都不同。这些发现表明,pBS582特有的序列分散在整个盘基网柄菌基因组中,在单倍体基因组中大约重复100次。Northern印迹分析显示,与pBS582 DNA杂交的RNA在生长和发育的所有阶段都存在,似乎不受发育调控。对其他黏菌(巨大盘基网柄菌、紫色盘基网柄菌和紫色多孢黏菌)的DNA进行Southern印迹分析,以确定pBS582序列是否存在于其他黏菌物种中。观察到pBS582与两种盘基网柄菌的DNA杂交,但与紫色多孢黏菌的DNA不杂交。因此,有可能将特定序列的杂交作为一种生化工具来研究不同黏菌物种的亲缘关系及其分子分类学。